HPV is an epitheliotropic virus that infects keratinocytes of the pharynx. Viral proteins affect the regulation of the cell cycle and cell repair systems, which can induce transformation of normal cells into cancer cells. Patients with HPV-positive pharyngeal tumors are more responsive to treatment with radiochemotherapy than patients with HPV-negative pharyngeal tumors. Due to the severe side effects of the treatment, it is reasonable to reduce the intensity of treatment for patients with HPV-positive pharyngeal cancer. This includes reducing the radiation dose and lowering the concentration of chemotherapy or replacing used cytostatics with less harmful ones. We evaluated the response of HPV-positive (SCC090) and HPV-negative (Detroit 562) cell lines to radiation and exposure to chemotherapeutics cisplatin, bleomycin, and carboplatin. We used the clonogenic and metabolic activity assay to evaluate the effect of irradiation and exposure to chemotherapeutics cisplatin, bleomycin, and carboplatin. We have shown that the HPV-positive (SCC090) and HPV-negative (Detroit 562) cell lines are essentially equally sensitive to radiation and chemotherapeutics cisplatin, bleomycin, and carboplatin. We also demonstrated that chemotherapeutics bleomycin and carboplatin have the same effect on the SCC090 and Detroit 562 cell lines. Of the selected chemotherapeutics, cisplatin had the most significant effect on cell lines SCC090 and Detroit 562. For more detailed characterisation of SCC090 and Detroit 562 cell lines, further in vitro studies are required, which include monitoring of DNA damage repair and determination of cell distribution in the cell cycle phases after irradiation or exposure to chemotherapeutics. The effects of other chemotherapeutics on cell lines could also be compared. The comparison between the cell lines could also be performed in vivo, in laboratory mice.